Frictional trans-epithelial tissue disruption collection apparatus and method of inducing an immune response

ABSTRACT

The invention relates to trans-epithelial frictionally abrasive tissue sampling devices for performing biopsies and methods of inducing an immune response against a pathogen, wherein epithelial cells containing the pathogen are disrupted with the frictionally abrasive tissue sampling device to introduce the pathogen into the bloodstream of a patient.

PRIORITY CLAIM

This application is a continuation of (1) U.S. Utility application Ser.No. 16/371,027 filed Mar. 31, 2019, which is a divisional of (2) U.S.Utility application Ser. No. 15/603,374 filed May 23, 2017 which issuedas U.S. Pat. No. 10,258,780 on Apr. 16, 2019, which is a divisional of(3) U.S. Utility application Ser. No. 15/208,603 filed Jul. 12, 2016which issued as U.S. Pat. No. 9,687,642 on Jun. 27, 2017, which is acontinuation of (4) U.S. Utility application Ser. No. 14/602,002 filedJan. 21, 2015 which issued as U.S. Pat. No. 9,393,394 on Jul. 19, 2016and which is a divisional of (5) U.S. Utility application Ser. No.14/154,447 filed Jan. 14, 2014, abandoned, which is a divisional of (6)U.S. Utility application Ser. No. 12/669,638 filed Jan. 19, 2010, whichissued as U.S. Pat. No. 8,652,067 on Feb. 18, 2014, which is theNational Phase of (7) Patent Cooperation Treaty Application US08/70341filed Jul. 17, 2008, which claims priority to (8) U.S. ProvisionalApplication No. 60/950,280 filed Jul. 17, 2007. These applications(1)-(8) are herein expressly incorporated by reference in theirentireties and for all purposes.

FIELD OF THE INVENTION

The invention relates to epithelial tissue sampling and collectiondevices for performing biopsies from lesions and anatomical landmarks atrisk of neoplastic transformation, including but not limited to thesquamo-columnar junction of the female cervix, methods of inducing animmune response against a pathogen and methods of trans-epithelial drugdelivery.

DESCRIPTION OF THE RELATED ART

Previous devices include brushes with rigid bristles that puncture andshear epithelial surfaces (U.S. Pat. Nos. 5,535,756; 6,258,044;6,376,905; 6,494,845 and 6,132,421), single metal or plastic curettesthat extend in a parallel direction to the applicator handle and aremuch larger than the innovation (U.S. Pat. Nos. 4,641,662 and6,730,085), scalpels or similar bladed sharp cutting tools (U.S. Pat.Nos. 5,857,982; 5,800,362; 3,774,590; 5,092,345; 4,061,146; 5,868,668;6,053,877; 5,470,308; 7,137,956, 4,168,698 and 4,757,826; and U.S.Publication Nos. 2005/0059905 and 2007/0093727), or very largeelectrified metal loops used to produce excisional biopsies (U.S. Pat.Nos. 5,913,857 and 5,951,550). One device performs simultaneous brushcytology and scrape biopsy on structures with an organic duct (U.S. Pat.No. 5,535,756).

Human papillomaviruses (HPV) are responsible for many cutaneous andmucosal lesions. Some viral genotypes are considered to be the causalagents of cervical cancer. Natural genital HPV infection seems to bepoorly immunogenic because of its nonproductive and non-inflammatorycharacteristics and also because of mechanisms developed by the virus tocounteract the immune response.

SUMMARY OF THE INVENTION

A first aspect relates to a fabric for functionally abrading epithelialsurfaces including a backing material and a plurality of fenestratedloops attached to the backing material, the loops having sufficientflexibility and rigidity to frictionally abrade the epithelial surfaces,wherein the loops are about 3 mm to about 25 mm in length, wherein theloops have a short hook end, and wherein the distance from the top ofthe loop to the bottom of the hook is less than 50% of the length of theloop.

A second aspect relates to an apparatus for obtaining a histologicalsample including a handle, a platform at a distal end of the handle, anda fabric for functionally abrading epithelial surfaces including abacking material and a plurality of fenestrated loops attached to thebacking material (26).

A third aspect relates to a method of inducing an immune responseagainst a pathogen that normally evades the immune system includingdisrupting epithelial cells containing the pathogen with a frictionaltrans-epithelial tissue disruption apparatus, and thereby introducingthe pathogen, DNA fragments, proteins or antigenic material into thebloodstream of a patient to elicit an immune response.

A fourth aspect relates to a method of trans-epithelial drug deliveryincluding disrupting tissue with a trans-epithelial tissue disruptionapparatus and applying a drug to intra-epithelial and sub-epithelialspaces created by the disrupting tissue.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Apparatus for frictional trans-epithelial tissue disruption ofan epithelial flat surface;

FIG. 2(A). Apparatus for frictional trans-epithelial tissue disruptionof an epithelial-lined canal surface with tapered cone tip, Side view;FIG. 2(B). apparatus for frictional trans-epithelial tissue disruptionof an epithelial-lined canal surface with tapered cone tip, obliqueview; FIG. 2(C). apparatus for frictional trans-epithelial tissuedisruption of an epithelial-lined canal surface with tapered cone tip,top view;

FIG. 3(A). Schematic diagram showing a method of frictionaltrans-epithelial tissue disruption of a flat epithelial surface; or FIG.3(B). schematic diagram showing a method of frictional trans-epithelialtissue disruption of an epithelial surface of a canal or body cavity;

FIG. 4. Frictional trans-epithelial tissue disruptor with a motorized orvibratory handle used to spin or agitate the fenestrated loops;

FIG. 5. Schematic diagram of an apparatus with a detachable platformthat anchors fiber loops at a distal end of the handle;

FIG. 6(A). Schematic diagram of frictional trans-epithelial tissuedisruption showing representation of tissue with a squamous epitheliallined surface; FIG. 6(B). Schematic diagram of frictionaltrans-epithelial tissue disruption with application of the frictionalbiopsy device to the body surface; FIG. 6(C). Schematic diagram offrictional trans-epithelial tissue disruption where simultaneouspressure, agitational, and rotational force splays and separates thehooks/loops. Frictional abrasive forces create heat which buckles theepithelial surface; FIG. 6(D). Schematic diagram of frictionaltrans-epithelial tissue disruption where sufficient abrasion createsshearing and fracture of the epithelial surface at varying depths whichcould include fracture through the basement membrane into thesubcutaneous layer; FIG. 6(E). Schematic diagram of frictionaltrans-epithelial tissue disruption where the hooks insinuate into thefracture plane, and with additional abrasive forces continue to shearthe tissue fragments, while simultaneously retaining the tissue forcapture and collection; FIG. 6(F). Schematic diagram of frictionaltrans-epithelial tissue disruption at the completion of the biopsyprocess, the collection of hooks arranged in rows create channels whichcollect and sequester the tissue and cell cluster fragments within thechannels created in the device. When the device is removed from theepithelial surface, additional sample is captured and held due to theflexibility and recoil of the hooks;

FIG. 7(A). Side view of a focal biopsy apparatus, depicted at the outerlip of the cervix (exocervix); FIG. 7(B). Schematic diagram of apparatusfor focal biopsies with an enlarged view of the platform and loops;

FIG. 8(A). Side view of apparatus for simultaneous biopsy of epithelialsurfaces and canal-like surfaces. Longer central core fibers toinsinuate into a canal and a perimeter of approximately 3 mm fiberscontact an outer epithelial surface; FIG. 8(B). Schematic diagram ofapparatus for simultaneous biopsy of epithelial surfaces and canal-likesurfaces with enlarged view of platform and loops.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Definitions

The transitional term “comprising” is synonymous with “including,”“containing,” or “characterized by,” is inclusive or open-ended and doesnot exclude additional, unrecited elements or method steps.

The transitional phrase “consisting of” excludes any element, step, oringredient not specified in the claim, but does not exclude additionalcomponents or steps that are unrelated to the invention such asimpurities ordinarily associated therewith.

The transitional phrase “consisting essentially of” limits the scope ofa claim to the specified materials or steps and those that do notmaterially affect the basic and novel characteristic(s) of the claimedinvention.

As used herein, the term “fenestrated loop” refers to a hooked,“candy-cane” shape formed by severing a loop, wherein a short, hookedend is less than about 50% of the length of the loop. In someembodiments, a fenestrated loop is formed by severing a loop once,leaving a short arm adjacent to the fenestrated loop.

Focal Biopsy

Some embodiments relate to a trans-epithelial, frictional tissuesampling and collection device to perform biopsies of lesions suspectedof harboring disease. In some embodiments, a lesional biopsy site is nolarger than about 10 mm in diameter (i.e., focal biopsy). In someembodiments the lesions are accessible to an examiner during routineexamination. In other embodiments, the surface is accessible followingentry into a body cavity through a natural orifice or surgical channelvia a trochar and inspection using an endoscope with a biopsy port. Thedevice head remains on the lesion or area of intended biopsy/therapy dueto the rigid nature of the applicator. Referring to FIG. 7, a focalbiopsy apparatus is configured with loops that are about 3 mm to about25 mm in length, preferably about 3 mm in length, wherein the loops havea short hook end, wherein the distance from the top of the loop to thebottom of the hook is less than 50% of the length of the loop.

Regional Biopsy

In some embodiments, the intent is to biopsy and screen large geographicareas of tissue at risk for disease (e.g., neoplastic transformationsuch as, but not limited to, the squamo-columnar junction of the femalecervix in the presence or absence of visualized lesions). The deviceprovides samples of clumps or clusters of excavated epithelial tissuefragments for analysis, in contrast to other methods disclosed in priorart that provide surface and exfoliated cells by sweeping the cells fromsuch target tissue sites, commonly with blunt spatula or soft bristlebrush devices. The intent is to remove tissue based evidence withfrictional biopsy of the larger area, which may range from 10-40millimeters in diameter.

Simultaneous Biopsy of Epithelial Surfaces and Canal-Like Structures

In some embodiments, the device contains a central core of longerfenestrated loops (e.g., about 4-7 mm long), surrounded by a wider rimof shorter fenestrated loops (e.g., about 3 mm in length). The longerloops are geometrically suited to insinuate within a central canalstructure, such as the endocervical canal of the cervix. There issimultaneously uniform contact of the fenestrated loop fiberscircumferentially around the endocervical canal on the flat exocervicalsurface. With rotation and agitation in a back-and-forth motion, tissueis harvested within the fenestrated loop channels as described above.

In some embodiments, the device contains a central core of longerfenestrated loops (e.g., about 4-7 mm long), surrounded by a wider rimof shorter fenestrated loops (e.g., about 3 mm in length). The longerloops are geometrically suited to insinuate within a central canalstructure, such as the endocervical canal of the cervix. There issimultaneously uniform contact of the fenestrated loop fiberscircumferentially around the endocervical canal on the flat exocervicalsurface. With rotation and agitation in a back-and-forth motion, tissueis harvested within the fenestrated loop channels as described above.

Frictional Tissue Sampling and Collection Biopsy Devices

The frictional tissue sampling and collection biopsy devices disclosedherein utilize a fabric that includes minute plastic (e.g., nylon) fiberloops that are fenestrated at a minimal distance from the apex of theloop. The loops flex but do not fracture under minimal to moderate forceor separate under pressure.

The semi-rigid loops may be pressed in a rotational manner (e.g., insweeping or circular motion) away from or toward the operator,perpendicular, or at an angle into epithelial tissue surfaces. Thesemi-rigid loops remain flexible enough to cause separation of thefenestrated ends, creating frictional forces sufficient to cause localheating and buckling of the epithelial surface away from the underlyingstroma. The loops are fenestrated such that with applied pressure theyare flexible enough to open and provide access to a “collection well”for histological fragments. The tips of the fiber hooks are orientedaway from the tissue. On pressing and rotation across the tissuesurface, the fibers scrape, buckle and shear the epithelium from theunderlying stroma. The fragments are excoriated from the tissue surfacethrough the concomitant application of frictional forces applied to thetissue surfaces by the fenestrated loops. The frictional forces overcomethe adhesive and binding forces of the tissue below to release fragmentsof various shapes and size, all eligible for collection in a histologylab, and subsequent processing and analysis.

The semi-rigid loops (e.g., made of nylon) hold the tissue fragmentsafter excoriation because the loops are elastic enough to sufficientlyre-close and capture the remove tissue. In addition, the spaces betweenthe fibers also retain excoriated tissue. The frictional forces exceedthe binding forces afforded by adhesion molecules which anchor epitheliato the basement membrane, as well as disrupting Van der Waals forces.

Once the epithelium is frictionally sheared from the underlying stroma,the tissue clumps and epithelial fragments are swept and excavated bythe distal most curved apex of the loop and entrapped within thegeometrically suited spaces between the closed, fenestrated loops. Thus,the method is frictional abrasion, excavation via rotation and otherdirectional motion, and tissue collection within inter-loop channels.

The fabric can be cut into uniform shapes such as a circular disc orstraight edge shape(s) and with uniform height, allowing the device toprovide 360 degree coverage of tissue surfaces over suspected lesions,without a gap within the circumference of the device. This is indistinction to bristle brushes which are spiral or bent in shape, whichpresent surface gaps that do not allow uniforms contact with the targettissue, and gaps that cause migration of the device from the lesion sitetoward the direction of rotation when such devices are pressed ontolesions and rotated or moved for tissue harvesting.

Following biopsy, the fabric, fibers, and/or device head (all with thetissue entrapped between the fibers) are removed and placed in a vial ofliquid fixative for later laboratory work. A laboratory may remove thetissue from the device and process it for analysis. Therefore, one mayintentionally design the device in an embodiment in which the user couldeasily decouple the device head from the device shaft. For example, someembodiments may have the shaft inserted into the head via a clip orscrew thread mechanism, a key-in-lock design with a pressure releasebutton, or a luer-lock type of attachment. Once the biopsy is obtained,the head and handle/shaft parts can be de-coupled, wherein the handlecan be discarded, or sterilized and re-used, and the head immersed in avial of fixative.

Some methods for removal of tissue from the fiber assembly includerinsing under pressure, immersion and agitation manually ormechanically, or by sonication. Alternatively, the fibers can be shearedfrom the fabric on telfa or other filter paper, and the fibers pluckedoff the paper leaving the entire biopsy specimen. Alternatively, aftertissue is collected into the device channels, tissue may deposited viarotation or agitation in a vial of liquid fixative, rinsed off thedevice under pressurized spraying, or removed from the nylon fibers bycutting away the nylon fibers from the fabric (e.g., onto filter paper),thus leaving the tissue on the paper, which can be immersed in fixative.

In preferred embodiments, the fabric fibers are manufactured in asimilar manner to Velcro® or other hook and pile type fastener, wherestrands are longer than conventional hook and pile, about 3 mm inlength, fenestrated closer to the apex of the loop instead of close tothe base of one arm of the loop, and thus appear V-wishbone shaped. Theyhave a short hook end with the curvature starting at 2 mm from the base.Because the loop strands are longer, they flex and bend to a greaterangle and twist with greater elasticity when rotated or agitated whencompared with standard Velcro® Because the fenestration is closer to thebase in standard Velcro®, the loops fenestrations are unlikely toseparate, leaving the curved smooth surface of the loop in contact withthe tissue, not providing sufficient frictional forces during rotationto shear and separate the epithelium form the underlying basementmembrane and stroma.

Preferred embodiments utilize minute plastic fenestrated loops that arepressed perpendicular or at an angle into epithelial tissue surfaceswhich, upon rotational or agitational pressure forces, cause tissueepithelial fragments to be frictionally separated from the underlyingtissue basement membrane and stroma. The channels between thefenestrated loops entrap and collect the tissue fragments. The processis similar to curettage with a blunt curved tool, which also scrapes,shears and strips epithelium from the underlying stroma of targettissues. On the other hand, the process is in contrast to sharpcurettage where the purposefully sharp edge of the curette firstincises, pierces, then shaves and scoops epithelium and underlyingstroma from the tissue surface. The process described herein is lessperceptible to patients than conventional biopsies and causes a smalleramount of blood loss and trauma.

In one aspect, the present invention relates to a frictionaltrans-epithelial tissue apparatus. In some embodiments, the apparatuscomprises 3 mm or smaller loops adherent to and projecting perpendicularfrom a platform, with a density of 5-50 loops per square inch, evenlyspaced or arranged in rows. The loops may be intact or fenestrated atthe center or at their lateral aspect to allow for added flexibility andconstructed from plastic, metal, or another stiff material. The roundedend of the loop is opposite the platform.

Loops of sufficient flexibility to withstand frictional forces and notfracture, and of sufficient tensile strength to generate sufficientfrictional shear force during a sweeping or circular motion of thedevice to remove epithelium from tissue. The space between loops mayserve to capture and harbor the sampled tissue.

In some embodiments designed for focal lesional biopsy, a flat, flexibleplatform, which anchors the loops may be of any size, but is mostpractically approximately 5-10 mm in diameter and circular in shape. Theshape may be another geometrical design if it affords an advantage incovering the target tissue area for sampling. The platform may be hingedin such a way that it can be folded or compressed, inserted through asmall endoscopic channel, and then reinstated to its original state as aplatform with a sampling surface. It may be comprised of plastic, cloth,or another composite material. The loops are threaded through andproject away from the platform towards the tissue surface. Someembodiments may comprise a hub fiber or “pin” that penetrates andanchors the center of the disc on the target biopsy area, serving as acentral post to rotate the disc around for stability.

In other embodiments intended to screen a larger, regional tissue siteat risk for neoplastic transformation or other disease process, theoptimal shape is circular, the diameter could range from about 10-50 mm,and the loops project at varied distances from the platform to towardsthe tissue surface. For the purpose of histological screening to detectcervical neoplasia, the central 5 mm diameter disc projects longer (5-25mm) fenestrated loop fibers, and is surrounded circumferentially by theaforementioned approximately 3-23 mm long loop fibers. The longer fibersinsinuate inside canal structures, (e.g., the endocervical canal)simultaneously with contact of the shorter fibers with an outerendothelial surface (e.g., the exocervical surface). Upon pressure androtation or agitation, the endocervical and exocervical tissues can besimultaneously frictionally sheared and collected. Histologicalscreening may be necessary to correctly reflect the presence or absenceof epithelial pathology, because adhesion molecules may preventrepresentative exfoliation from diseased tissue in some cases, leavingcytological screening methods lacking in accuracy.

Preferably, a frictional trans-epithelial biopsy sample is taken from alesion or an anatomical region that is predisposed to disease.

Some embodiments comprise a plastic, metal, or mixed compositioncylinder or curved convex head, which provides a flat surface for theplatform to be attached to. It is equal or greater in diameter to theplatform. The cylinder is 5-10 mm in length while the flat or convexbase is less than about 3 mm thick.

Some embodiments comprise a rod or cylindrical shaped applicator probecomprised of any suitable material (e.g., wood, plastic, paper ormetal), which has the base, platform and loop unit at its most distalend, wherein the applicator probe is approximately 2-5 mm in diameterand 15-30 cm in length. It is constructed larger or smaller depending onthe access to the tissue surface. The shaft of the rod or cylindricalshaped applicator probe may be rigid or semi-rigid so as to not bow orarc when pressure is transmitted from the handle to the device head.

A handle into which the applicator probe can be transfixed is optionallymechanical, providing motorized rotational, drill-like movement oragitating vibration.

The device handle will be composed of stiff material, preferably plasticsimilar to Lucite, clear or opaque in coloration, rigid nylon plastic,or alternatively could be wood or metal. The device head can take mayshapes, cylindrical or tapered in design, but the distal most platformface is circular, square, or polygonal, and may be composed of plastic,(e.g., nylon). The diameter may range from 5-50 mm. The fabric is weldedto the nylon platform ultrasonically, or may alternatively be attachedvia adhesive, or via a rim or collar (e.g., which snaps on to theplatform into a recess in the head of the device).

In some embodiments, the operator examines tissue surfaces and choosesan area to be sampled based on the presence of a suspicious lesion. Inother embodiments, the operator chooses an anatomical landmark known tobe “at risk” for neoplastic or disease transformation for the purposesof sampling the entire chosen surface.

The handle or applicator probe is grasped at its proximal end or handle.

The distal portion or head of the device that contains the base,platform and loops that project perpendicular from the base towards thetissue surface with the more rounded ends that are pressed against thetissue surface.

With moderate pressure, the examiner simultaneously presses and rotatesthe device against the tissue several times in a clockwise orcounterclockwise direction, opening or separating the fenestrated loops,thus performing frictional disruption of the tissue surface.Alternatively, a sweeping motion may be used. If a motorized handle isused, it can be activated to assist in the rotation or vibration of thedevice.

The harvested tissue is collected from the tissue surface, and sometissue already trapped in the loops themselves is inspected and can beteased from the loops, or the loops transected from the fabric andseparated, and the remaining tissue placed in a fixative solution.

As shown in FIG. 1, fabric with fenestrated loops (1) is connected toplatform (2), which is in communication with head (3), located at adistal end of handle (5), optionally including an elongated rod (4).Referring to FIG. 3A, moderate force (8) is applied against a tissuesurface (7). The device head is rotated (9) on the surface tofrictionally separate or agitate the surface epithelium. The device headis rinsed or placed with tissue in the loops into fixative forsubsequent pathological analysis.

An apparatus with a conical platform is depicted in FIG. 2. In FIG. 2A,fabric with fenestrated loops (1) is connected to conical platform (6).Referring to FIG. 3B, an apparatus with a conical platform may beinserted into a canal or cavity. The device head is rotated (9) whilemaintaining pressure force in direction (8). The device head with tissuein the loops is rinsed or placed into pathological fixative.

An apparatus with a motor configured to rotate the platform is depictedin FIG. 4. Fabric with fenestrated loops (1) is attached to platform (2)on head (3) at the distal end of an elongated rod (4), which is attachedto a motorized handle (5).

In some embodiments, the head is detachable from the elongatedrod/handle. Referring to FIG. 5, a detachable head configuration allowsthe distal portion with head (3), platform (2), together with attachedfabric containing loops, to be detached and placed into a preservativemedium for later tissue removal and pathological processing. Someembodiments may have the shaft inserted into the head via a clip orscrew thread mechanism, or a luer-lock type of attachment (23). Tissuefragments that remain attached to the detachable head are in addition toany free tissue obtained and collected from the tissue surface or thedevice as a result of the frictional tissue sampling.

Referring to FIG. 6, epithelial tissue samples are obtained byfrictional trans-epithelial tissue disruption. A representation oftissue with a squamous epithelial lined surface is depicted in panel(A). The squamous epithelial multilayer (11) is shown with superficialflat and basal cuboidal epithelium. Basement membrane (12) separates thesquamous epithelial multilayer from the subcutaneous tissue stroma (13)and the underlying sub-stromal tissue (14). FIG. 6B depicts applicationof the frictional biopsy device to the tissue surface. The device head(3) is applied (24) to a chosen area where curved portions of thefenestrated loops (1) press against the epithelial surface. Arepresentation of two abutting hooks is shown, creating a collectionchannel. A shorter arm (15), adjacent to the fenestrated loop (1), mayremain following severing of an initial continuous loop to create thefenestrated loop. In FIG. 6C, simultaneous pressure, agitational, androtational force (16) splays and separates the hooks/loops. Frictionalabrasive forces create heat which buckles the epithelial surface.Referring to FIG. 6D, sufficient abrasion creates shearing and fractureof the epithelial surface at varying depths which could include fracturethrough the basement membrane into the subcutaneous layer. As shown inFIG. 6E, the hooks insinuate into the fracture plane, and withadditional abrasive forces continue to shear the tissue fragments, whilesimultaneously retaining the tissue for capture and collection. At thecompletion of the biopsy process (FIG. 6F), the collection of hooksarranged in rows creates channels that collect and sequester the tissueand cell cluster fragments within the channels. When the device isremoved from the epithelial surface, additional sample collection isachieved due to the flexibility and recoil of the hooks.

Referring to FIG. 7A, frictional trans-epithelial tissue disruption witha focal biopsy apparatus is shown at the outer lip of the exocervix(17), alternatively known as the “transformation zone” of the cervix(18). In this configuration, fenestrated loops (1) approximately 3 mm inlength are used to disrupt and collect tissue fragments. FIG. 7B depictsan enlarged focal biopsy apparatus, with an enlarged view of fenestratedloops (1) attached to platform (2).

Referring to FIG. 8A, simultaneous trans-epithelial biopsy of epithelialsurfaces and canal-like surfaces, in particular, biopsy of theendocervical canal (20) and the exocervical area around the endocervicalcanal (i.e., the transformation zone) is shown (19). Referring to FIG.8B, a central core of elongated loops of about 5-25 mm in length (21)are surrounded by a wider rim of shorter fenestrated loops of about 3-23mm in length (22).

The frictional tissue sampling and collection device can be used on anybody surface, both external to the body, body cavities, or on internalorgans. To access epithelial surfaces of internal body organs, thedevice head may be deflated, folded or collapsed to pass through a smallaperture or port, and re-opened or expanded to fully expose the fabricto the biopsy surface. This device can be used on humans or any otherliving organism with an epithelial surface. Any tissue surface may besampled. The ease of use in each case will be related to the strength ofthe individual tissue adhesion and binding forces in specific locations.The loops themselves can harvest the tissue and also serve as tissuecollection reservoirs for later storage once placed in a fixativemedium. The platform with the loops may be detached from any applicatorfor later examination and processing (i.e., decoupled from theinstrument used to press against tissue surfaces to obtain the tissuesample).

If the tissue surface is a canal or concave shaped area of the body,instead of a perpendicular platform design, the loops are directlyattached to the probe itself which are gradually tapered at the end tofacilitate insertion into the canal. The loops project perpendicularlyfrom the probe surface at its distal end, and the unit, once placed intothe canal that is lined on its surface with epithelium, contacts suchepithelium snugly.

The loops can be mounted on the platform or project from the rim surfaceof the platform, perpendicular or at an angle to the platform along themargin of the platform, or attached to other delivery applicators,including the examiner's gloved finger, or other surgical instruments.The platform can be any shape or size which can fit on a tissue surface.The base assembly can be any shape or size, and may be permanently rigidor collapsible.

If the tissue surface lies within a canal shaped tissue surface, theloops can be attached directly to the applicator probe, which can beinserted into the canal shaped body cavity. The probe with the loopsprojecting from the surface and contacting the epithelium is rotatedcausing the frictional disruption sampling from the tissue surface. Theshape of the probe can be constructed in any shape that allows a snugfit into the canal. The loops may be arranged in rows or equally spaced,allowing for maximal contact and tissue collection.

Some embodiments of the invention comprise a motorized mechanicalassistance via a mechanical handle into which the most proximal end ofthe applicator probe is inserted. Such mechanical assistance may enhancethe rotational or vibratory force that the device transmits to thetissue after contact is established. This can increase the frictionalforces and the speed of the tissue disruption/sampling and shorten theprocedure time.

Preferred Parameters of Fibers

The frictional sampling loops of the invention are collectively referredto as fenestrated loop fibers. In particularly preferred embodiments,the fibers are made using the hooked side of a modified Velcro® or otherhook and pile type fastener, where the strands are about 3 mm in lengthand are V-wishbone shaped. They have a short hook end (25) with thecurvature starting at 2 mm from the base. In various embodiments, theloops may be 2.5-25 mm in length, 3-5 mm in length, 3-10 mm in length,3-15 mm in length, 3-20 mm in length or 3-25 mm in length.

In comparison, standard Velcro® is about 2 mm long and is more hooked.Thus, the loops of the present invention are longer than those ofstandard Velcro®, they are made of a similar nylon material comparedwith standard Velcro®, are more flexible when rubbed on a tissue surfacedue to their length, and they have shorter loops that hook nearer to theend of the strands. In particular, the distance from the top of the loopto the bottom of the hook is preferably less than 50% of the length ofthe loop, more preferably less than 40%, still more preferably less than30%, and even more preferably less than 20% the length of the loop. Thisdistance is also preferably at least 1% the length of the loop, morepreferably at least 5% the length of the loop, and still more preferablyat least 10% the length of the loop. Thus, the invention includes hooksin all of the ranges between any of the preferred minimum distances andany of the preferred maximum distances. The bottoms of the hooks arepreferably arranged so that they are all approximately the same distancefrom the loop, although this is not strictly necessary. Because thehooks are cut at a relatively distal location, the ends of the hooks aremore accessible to the tissue surface allowing for uniform transmissionof frictional forces to the tissue surface. As a result, the action ofthe fibers more effectively buckle and shear the tissue, while the loopssweep over and capture the tissue.

In a preferred embodiment, the loop fibers are arranged so as toefficiently capture tissue. Thus, in one preferred embodiment, thefibers are arranged in an orderly orientation. For example, the fiberscan be arranged in rows between which the tissue can be captured. Thehooks can be arranged to be oriented at approximately the same angle anddirection in each of the fibers. Thus, the fibers can be organized suchthey all have a consistent direction and angle of orientation. Inaddition, the spacing between each of the fibers can be made to be thesame or different.

In use, the device can be oriented so that the fibers are perpendicularto tissue, and then pressure is applied. As a result, the epithelialsurface is frictionally sheared. Thus, the fibers are preferably mountedon a flat or curved platform, optimally 4-10 mm in diameter so asoptimize this process. However, alternatively shaped platforms can alsobe used in certain embodiments. Because the fibers can be mounteddirectly on the platform, which may be flat or slightly curved, theorientation remains evenly spaced and the spaces inside the fenestratedloops and between them remain evenly distributed to facilitate tissuecapture.

In some embodiments the platform may in the form of a thumbtack, whereinit is attached to the handle. However, the platform and handle may takeon a variety of forms. It is envisioned that the handle and the platformmay be molded as one piece, and the fibers (e.g., modified Velcro®) maybe attached with adhesive or via ultrasonic or thermal welding of thefabric to the platform.

Method of Inducing an Immune Response by Autoinoculation

In some embodiments, the trans-epithelial, frictional tissue samplingand collection devices described herein are utilized to agitate anddisrupt epithelial cells containing a pathogen, or cellular proteinsaltered by a pathogen, to induce an immune response against thepathogen. This results in auto-inoculation of tissues that harborpathogens and macromolecules such as virally altered DNA and/oroncogenic proteins. The method may also be termed therapeutic frictionalabrasion-excoriation. This method is advantageous when a pathogen isnormally able to evade an immune response. For example, some virusesremain in surface epithelial layers where they are sequestered from theimmune system. Other viruses may be integrated into cellular DNA,thereby evading immune detection.

The methods of inducing an immune response against a pathogen thatnormally evades the immune system comprise the steps of a) disruptingepithelial cells containing the pathogen, virally altered DNA, orcellular oncoproteins with a micro-curettage device described herein,and b) introducing the pathogen into the bloodstream of a patient toelicit an immune response.

In some embodiments, the trans-epithelial, frictional tissue samplingand collection devices described herein are utilized to disruptepithelial cells to induce an immune response against humanpapillomaviruses (HPVs). HPVs are persistent viruses that can remain intheir hosts for long periods of time before causing any ill effects.Generally, the host reacts to viral pathogens by generating both humoraland cell-mediated responses. Humoral responses are typicallyantibody-mediated and involve the secretion of antibodies such asimmunoglobulin A (IgA) and immunoglobulin G (IgG) by B lymphocytes.Cell-mediated responses, on the other hand, are carried out by immuneeffector cells such as dendritic cells (DCs), natural killer (NK) cells,macrophages and T lymphocytes which secrete a number of cytokinesincluding interferons (INF) and tumor necrosis factor (TNF), andup-regulate the expression of Fas ligand (FasL) and TNF-relatedapoptosis inducing ligand (TRAIL) on their cell surface.

In the case of HPV infection, the immune response is frequently weak orundetectable, and accompanied by little or no inflammation. Even when animmune response is elicited, it may not be able to clear the virus.Disruption of the epithelial surface by frictional tissue disruptioninduces repair and inflammation and serves to auto-inoculate thepatient. Without wishing to be bound by any theory, exposure of theepithelial surface to frictional tissue disruption, uniquely induced bythe apparatus and methods disclosed herein through local heating fromfriction forces exerted, may enhance the induction of repair,inflammation and an immune response following patient autoinoculation.Agitation or scrubbing of a lesion serves to introduce viral particlesinto the bloodstream of a patient where they can trigger a humoral orantibody related immune response. In addition the method can fracturecells releasing antigens locally within the tissue stroma inducing acell mediated response associated with the release of cytokines andattraction of helper and killer T cells to the sampled tissue area.

Advantageously, the method of the present invention auto-inoculates apatient with viral particles of the specific viral serotype(s) that thepatient is infected with. In contrast, current vaccine strategies areeffective on a subset of HPV strains. For example, GARDASIL® by Merck &Co., Inc. is indicated to help prevent cervical cancer, precancerous andlow-grade cervical lesions, vulvar and vaginal pre-cancers and genitalwarts caused by human papillomavirus (HPV) types 6, 11, 16 and 18 andCervarix™ by GlaxoSmithKline is an HPV 16/18 cervical cancer candidatevaccine. The vaccine is commonly injected in a limb, not the targetorgan at risk, the cervix, and has been only documented to elicit ahumoral antibody immune reaction.

Drug Application

In some embodiments, an adjuvant drug or an immune modulating agent isused in combination with the autoinoculation method, thus augmenting animmune response. For example, Imiquimod (Aldara® topical cream,manufactured and marketed by Graceway Pharmaceutical Company) isapproved for the treatment of actinic keratosis, external genital wartsand superficial basal cell carcinoma (sBCC), a type of skin cancer. Animmune response may be enhanced by using such immune modulating agentsin combination with autoinoculation by the methods described herein. Theadjuvant drug can be applied to the fenestrated loop fibers directlyakin to toothpaste on a toothbrush, or a channel within the applicatorcan be used to transmit the drug from the top of the handle by means ofa squeeze bulb or syringe, through a small lumen in the center of thefabric disc, concomitant with the tissue disruption, delivering druginto the fracture crevices created during the frictional buckling andshearing process created by the device.

Some embodiments comprise a method of drug delivery to a pathologicallesion or areas of tissue that concomitantly disrupts tissue planes,creating crevices or pathways for drugs to enter via intra-epithelialand sub-epithelial spaces. This is in contrast to topical therapies,which are slowly absorbed into and through the epithelia. Intra-lesionalapplication is more focused and requires less drug, presenting less riskof side effects.

Any type of drug (e.g., ablative, antibiotic, antiseptic, immunemodulating, etc. may be used.

In some embodiments, drug is delivered via an applicator comprising afabric with fenestrated loops as described herein. Drug is applied in amanner akin to applying toothpaste to a toothbrush, or drug may injectedonto the platform or the apparatus via a channel leading through ahollow applicator handle. The drug application apparatus may optionallyhave an element through which the drug is delivered (e.g., a syringewith a locking mechanism). Drug is applied to a “wound” created byfrictionally agitating the tissue. In some embodiments, the fenestratedloops may be impregnated with a drug during manufacture, wherein thedrug leeches out into the disrupted tissue when the fiber contacts andmacerates/disrupts the tissue.

While the present invention has been described in some detail forpurposes of clarity and understanding, one skilled in the art willappreciate that various changes in form and detail can be made withoutdeparting from the true scope of the invention. All figures, tables, andappendices, as well as patents, applications, and publications, referredto above, are hereby incorporated by reference.

1. A device for shearing clumps of tissue comprising: a glove finger ora finger cot; a fabric comprising: a platform disposed on a distal endof the glove finger or the distal end of the finger cot; and a firstplurality of fenestrated loops attached to the platform, where the firstplurality of fenestrated loops each comprise: a first length; and afirst hook end, where a first distance from the top of the firstplurality of fenestrated loops to the bottom of the first hook end isless than approximately 20% of the first length.
 2. The device of claim1, the platform is flat.
 3. The device of claim 1, the platform isflexible.
 4. The device of claim 1, the platform is collapsible.
 5. Thedevice of claim 1, where the first plurality of fenestrated loops arearranged in rows.
 6. The device of claim 1, where the first plurality offenestrated loops are arranged equally spaced.
 7. The device of claim 1,where the first length is between: a lower limit of 2.5 mm; and an upperlimit of 25 mm.
 8. The device of claim 1, the platform is circular inshape.
 9. The device of claim 8, the platform is between: a lower limitof 5 mm in diameter; and an upper limit of 10 mm in diameter.
 10. Thedevice of claim 1, the platform is circular in shape.
 11. The device ofclaim 1, the platform is straight edged in shape.
 12. The device ofclaim 1, further comprising a second plurality of fenestrated loopsattached to the platform, where the second plurality of fenestratedloops comprise: a second length; and a second hook end, where a seconddistance from the top of the second plurality of fenestrated loops tothe bottom of the second hook end is less than approximately 20% of thesecond length.
 13. The device of claim 1, where the clumps of tissuecomprise a histological tissue sample.
 14. A method for obtaining ahistological tissue sample comprising: (a) placing a finger cot on afinger of a gloved hand, the finger cot comprising: a fabric comprising:a platform disposed on a distal end of the finger cot; a plurality offenestrated loops attached to the platform, where the plurality offenestrated loops each comprise: a length; a hook end, where a distancefrom the top of the plurality of fenestrated loops to the bottom of thehook end is less than approximately 20% of the length; (b) applying thefinger cot to an tissue surface; (c) applying a moderate force to thefinger cot, where the moderate force causes one or more hook ends of theplurality of fenestrated loops to flex but not to shear or separate anepithelium from an underlying stroma; and (d) applying a frictionalforce to the finger cot, where the frictional force is acting on the oneor more hook ends to recoil against the tissue surface to removetransepithelial cells from the underlying stroma.
 15. The method ofclaim 14, where the frictional force is acting to splay and separate oneor more of the plurality of fenestrated loops.
 16. The method of claim14, where the frictional force causes the one or more hook ends toabrade the tissue surface leading to tissue disruption.
 17. The methodof claim 14, where the finger is an index finger.
 18. A method forshearing clumps of tissue from a wound surface comprising: (a) placing afinger cot on a finger of a gloved hand, the finger cot comprising: afabric comprising: a platform disposed on a distal end of the fingercot; a plurality of fenestrated loops attached to a backing material,where the plurality of fenestrated loops each comprise: a length; a hookend, where a distance from the top of the plurality of fenestrated loopsto the bottom of the hook end is less than approximately 20% of thelength; (b) applying the finger cot to a wound tissue surface; and (c)applying a frictional force to the finger cot, where the frictionalforce is acting on one or more hook ends of the plurality of fenestratedloops, where the one or more hook ends react to the frictional force byrecoiling against the wound tissue surface to shear clumps of tissuefrom an underlying stroma.
 19. The method of claim 18, where thefrictional force is acting to splay and separate one or more of theplurality of fenestrated loops.
 20. The method of claim 18, where thefrictional force causes the one or more hook ends to abrade the woundtissue surface leading to tissue disruption.